Development of a nucleotide prodrug of 4'-fluorouridine to improve tolerability

4'-fluorouridine (4'-FlU; EIDD-2749), a previously described broad-spectrum pyrimidine derivative, is orally efficacious against numerous high consequence viruses, including influenza virus, respiratory syncytial virus, and chikungunya virus. During the course of development of EIDD-2749, adverse gastrointestinal effects and weight loss were observed in mice at higher doses, presumably due to the rapid conversion to the active 5'-triphosphate in the gut. A synthetic campaign was launched to identify a prodrug derivative of EIDD-2749 with improved tolerability and sustained efficacy. A total of 15 monophosphate prodrugs were synthesized and assessed in mice for tolerability improvements and efficacy against a lethal challenge of influenza A/California/07/2009 (H1N1). From this library, EIDD-2838 stood out as meeting the desired criteria. Tissue distribution studies with EIDD-2838 revealed lower overall exposure to the active 5'-triphosphate when dosed with the prodrug, including a 4-fold reduction in the gut. Resolution of the phosphorous diastereomers of EIDD-2838 using a synthetic, copper-mediated chiral auxiliary approach revealed a single diastereomer, EIDD-3639, the P_S diastereomer of EIDD-2838, with retained efficacy in a low-dose (2 mg/kg) trial against H1N1 infection. An alternative route toward multi-gram scale preparation of EIDD-3639 was developed, utilizing a stereoselective chiral catalyst coupling reaction combined with a p-nitrobenzoate-assisted crystallization.